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pAC-LYC vector (V014693) Gene synthesis in pAC-LYC backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V014693 pAC-LYC In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pAC-LYC
Antibiotic Resistance:
Chloramphenicol
Length:
9476 bp
Type:
Gene cloning
Replication origin:
p15A ori
Host:
E. coli
Growth Strain(s):
Top10
Growth Temperature:
30℃

pAC-LYC vector Map

Copy Sequence View Map
pAC-LYC9476 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200cat promoterp15A oritet promotercrtEcrtIcrtBCmR

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pAC-LYC vector Sequence

LOCUS       Exported                9476 bp DNA     circular SYN 21-JUL-2025
DEFINITION  Exported.
ACCESSION   V014693
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9476)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9476)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9476)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1505..1533
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1993..2913
                     /gene="crtE"
                     /product="Geranylgeranyl diphosphate synthase"
                     /label=crtE
                     /label=Geranylgeranyl diphosphate synthase
                     /note="Geranylgeranyl diphosphate synthase from
                     Pseudescherichia vulneris. Accession#: P22873"
     CDS             4178..5635
                     /codon_start=1
                     /gene="crtI"
                     /product="Phytoene desaturase (lycopene-forming)"
                     /label=crtI
                     /label=Phytoene desaturase (lycopene-forming)
                     /note="Phytoene desaturase (lycopene-forming) from 
                     Pseudescherichia vulneris. Accession#: P22871"
                     /translation="MKKTVVIGAGFGGLALAIRLQAAGIPTVLLEQRDKPGGRAYVWHD
                     QGFTFDAGPTVITDPTALEALFTLAGRRMEDYVRLLPVKPFYRLCWESGKTLDYANDSA
                     ELEAQITQFNPRDVEGYRRFLAYSQAVFQEGYLRLGSVPFLSFRDMLRAGPQLLKLQAW
                     QSVYQSVSRFIEDEHLRQAFSFHSLLVGGNPFTTSSIYTLIHALEREWGVWFPEGGTGA
                     LVNGMVKLFTDLGGEIELNARVEELVVADNRVSQVRLADGRIFDTDAVASNADVVNTYK
                     KLLGHHPVGQKRAAALERKSMSNSLFVLYFGLNQPHSQLAHHTICFGPRYRELIDEIFT
                     GSALADDFSLYLHSPCVTDPSLAPPGCASFYVLAPVPHLGNAPLDWAQEGPKLRDRIFD
                     YLEERYMPGLRSQLVTQRIFTPADFHDTLDAHLGSAFSIEPLLTQSAWFRPHNRDSDIA
                     NLYLVGAGTHPGAGIPGVVASAKATASL"
     CDS             5653..6582
                     /codon_start=1
                     /label=crtB
                     /translation="MSQPPLLDHATQTMANGSKSFATAAKLFDPATRRSVLMLYTWCRH
                     CDDVIDDQTHGFASEAAAEEEATQRLARLRTLTLAAFEGAEMQDPAFAAFQEVALTHGI
                     TPRMALDHLDGFAMDVAQTRYVTFEDTLRYCYHVAGVVGLMMARVMGVRDERVLDRACD
                     LGLAFQLTNIARDIIDDAAIDRCYLPAEWLQDAGLTPENYAARENRAALARVAERLIDA
                     AEPYYISSQAGLHDLPPRCAWAIATARSVYREIGIKVKAAGGSAWDRRQHTSKGEKIAM
                     LMAAPGQVIRAKTTRVTPRPAGLWQRPV"
     CDS             complement(join(9039..9476,1..219))
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"